基于纳米免疫磁珠快速富集4种海洋致病性弧菌的研究
发布时间：2018-12-18 13:59
【摘要】：为研究海洋中常见致病性弧菌的快速分离富集方法——免疫磁珠法,本文以副溶血弧菌、哈维氏弧菌、创伤弧菌、灿烂弧菌为研究对象,制备纳米免疫磁珠,并通过一系列优化条件,用3M试纸计算捕获率来确定免疫磁珠最佳捕获条件。最后分别对4种致病性弧菌的不同菌液稀释浓度、对1株目标菌和8株非目标菌、4种混合菌进行捕获,从而评价免疫磁珠的灵敏度、特异性以及抗干扰能力。结果表明,4种磁珠偶联最佳抗体量为300、600、150、150μg。最佳捕获条件:副溶血弧菌免疫磁珠用量3mg,免疫反应45min,磁分离3min,p H 7.4缓冲液体系。哈维氏弧菌免疫磁珠用量3mg,免疫反应45min,磁分离1min,p H 6.0缓冲液体系。创伤弧菌免疫磁珠用量12mg,免疫反应60min,磁分离5min,p H 8.0缓冲液体系。灿烂弧菌免疫磁珠用量6mg,免疫反应30min,磁分离3min,p H 7.4缓冲液体系。在纯培养情况下,4种免疫磁珠灵敏度均达到10 CFU/m L数量级;分别对9株菌进行捕获,对目标菌的捕获率均达到87%以上,对非目标菌的捕获率均低于32%;对4种混合菌进行捕获时,免疫磁珠捕获率不受其它杂菌干扰。本研究证明了该免疫磁珠有较好的灵敏度、特异性和抗干扰能力,且与其它增菌方法相比,大大缩短了时间,具有较好的应用前景。
[Abstract]:In order to study the rapid isolation and enrichment of vibrio in the sea-immunomagnetic beads were prepared by using Vibrio parahaemolyticus, Vibrio Harvey's, Vibrio vulnificus and Vibrio splendens as research objects. Through a series of optimization conditions, the optimal capture conditions of immune beads were determined by calculating the capture rate of the 3M test paper. Finally, the dilution concentration of four pathogenic Vibrio vibrio strains in different liquid, one target bacterium, eight non-target bacteria and four mixed bacteria were captured to evaluate the sensitivity, specificity and anti-interference ability of the immune beads. The results showed that the best antibody concentration of four magnetic beads was 300600150150 渭 g. The optimal trapping conditions were as follows: the amount of immunomagnetic beads of Vibrio parahaemolyticus was 3 mg, the immune reaction was 45 min, and the magnetic separation was 3 min / h 7.4 buffer system. The dosage of immunomagnetic beads of Vibrio Harvey was 3 mg, the immune reaction was 45 min, and the magnetic separation was 1 min with pH 6.0 buffer system. The dosage of immunomagnetic beads of Vibrio vulnificus was 12 mg, the immune reaction was 60 min, and the magnetic separation was 5 min with pH 8.0 buffer system. The dosage of immunomagnetic beads of Vibrio splendens was 6 mg, the immune reaction was 30 min, and the magnetic separation was 3 mins / h 7.4 buffer system. In pure culture, the sensitivity of the four kinds of immunomagnetic beads was 10 CFU/m / L, 9 strains were captured, the capture rate of the target bacteria was more than 87%, and the capture rate of non-target bacteria was lower than 32%. The capture rate of immune beads was not disturbed by other hybrids when the four mixed bacteria were captured. This study proved that the immunomagnetic beads have better sensitivity, specificity and anti-interference ability, and compared with other methods of bacteriostasis, the time is greatly shortened, and has a better application prospects.
【作者单位】： 宁波大学海洋学院;
【基金】：海洋公益性行业科研专项;201105007号 宁波大学研究生科研创新基金项目;G14045号
【分类号】：R155.5
本文编号：2385949
[Abstract]:In order to study the rapid isolation and enrichment of vibrio in the sea-immunomagnetic beads were prepared by using Vibrio parahaemolyticus, Vibrio Harvey's, Vibrio vulnificus and Vibrio splendens as research objects. Through a series of optimization conditions, the optimal capture conditions of immune beads were determined by calculating the capture rate of the 3M test paper. Finally, the dilution concentration of four pathogenic Vibrio vibrio strains in different liquid, one target bacterium, eight non-target bacteria and four mixed bacteria were captured to evaluate the sensitivity, specificity and anti-interference ability of the immune beads. The results showed that the best antibody concentration of four magnetic beads was 300600150150 渭 g. The optimal trapping conditions were as follows: the amount of immunomagnetic beads of Vibrio parahaemolyticus was 3 mg, the immune reaction was 45 min, and the magnetic separation was 3 min / h 7.4 buffer system. The dosage of immunomagnetic beads of Vibrio Harvey was 3 mg, the immune reaction was 45 min, and the magnetic separation was 1 min with pH 6.0 buffer system. The dosage of immunomagnetic beads of Vibrio vulnificus was 12 mg, the immune reaction was 60 min, and the magnetic separation was 5 min with pH 8.0 buffer system. The dosage of immunomagnetic beads of Vibrio splendens was 6 mg, the immune reaction was 30 min, and the magnetic separation was 3 mins / h 7.4 buffer system. In pure culture, the sensitivity of the four kinds of immunomagnetic beads was 10 CFU/m / L, 9 strains were captured, the capture rate of the target bacteria was more than 87%, and the capture rate of non-target bacteria was lower than 32%. The capture rate of immune beads was not disturbed by other hybrids when the four mixed bacteria were captured. This study proved that the immunomagnetic beads have better sensitivity, specificity and anti-interference ability, and compared with other methods of bacteriostasis, the time is greatly shortened, and has a better application prospects.
【作者单位】： 宁波大学海洋学院;
【基金】：海洋公益性行业科研专项;201105007号 宁波大学研究生科研创新基金项目;G14045号
【分类号】：R155.5
【相似文献】
相关期刊论文 前1条
1 徐礼源;谢秀红;吴武鑫;谷康定;;免疫磁珠检测水中f2噬菌体方法的研究[J];中国卫生检验杂志;2008年04期
相关博士学位论文 前1条
1 覃昱;应用免疫磁珠分离及LAMP技术快速检测配方奶粉中克罗诺杆菌[D];南方医科大学;2014年
相关硕士学位论文 前1条
1 王四全;水中病毒膜吸附洗脱和免疫磁珠吸附法研究[D];华中科技大学;2009年
，本文编号：2385949
本文链接：https://www.wllwen.com/yixuelunwen/yufangyixuelunwen/2385949.html
